骨保护素和核因子κB受体活化因子配基对体外培养鸭破骨细胞的影响

Influnce of OPG and RANKL on duck′s osteoclasts formation and activation in vitro

探讨了核因子κB受体活化因子配基(Receptor activator of NF-κB ligand,RANKL)和不同浓度骨保护素(Os-teoprotegerin,OPG)对鸭破骨细胞(Osteoclasts,OC)生成和活化的影响。从7日龄内番鸭长骨骨髓分离破骨细胞,添加1,25-(OH)2D3进行诱导培养15 h后,更换含不同细胞因子的培养液(对照组:不加细胞因子;30μg/LsRANKL组;30μg/L sRANKL+10μg/L OPG组;10μg/L OPG组;50μg/L OPG组;100μg/L OPG组),继续培养。倒置显微镜进行OC形态学观察,比较各组培养3 d后抗酒石酸酸性磷酸酶(TRAP)阳性OC数量、培养7 d后扫描电镜观察象牙片吸收陷窝。结果表明:50μg/L OPG组、100μg/L OPG组OC数均极显著少于10μg/LOPG组(P<0.01);添加OPG组OC数均极显著少于30μg/L sRANKL组(P<0.01),象牙片吸收陷窝随OPG添加浓度的增加而减少,面积减小,陷窝深度变浅。RANKL能够促进OC的生成及活化,而OPG可与RANKL竞争性地结合RANK抑制OC的生成和活化,从而抑制OC的骨吸收。

The influnce of receptor activator of NF-κB ligand （RANKL） and different concentration of osteoprotegerin （OPG） on differentiation and activity of duck＇s osteoclasts was explored in the present experiment. Osteoclasts （OC） were isolated from long bone marrow of 1 to 7 days old＇ducks, and supplemented with 30 ng/ml sRANKL,30 μg/L sRANKL＋10 μg/L OPG, 10 μg/L OPG, 50 μg/L OPG and 100 μg/L OPG, respectively. The control group was supplemented nothing except a-MEM. The formation and activation of osteoclasts were detected （or examined） by ways of morphologic observation, histochemical staining for tartrate-resistant acid .phosphatase （TRAP） at the 3rd day and detection of lacunar resorption through scanning electron micrograph at the 7th day. Compared with groups supplemented with OPG, the number of TRAP positive osteoclasts in 30 μg/L sRANKL group increased very significantly （P〈0.01）. Compared with the 10 μg/L OPG group, the number of TRAP positive osteoclasts in 50 μg/L OPG and 100 μg/L OPG group decreased very significantly （P〈0.01）. The number and area of lacunar resorption decreased with concentration increasing of OPG. RANKL may advance OC formation and activation,while OPG may inhibits bone absorbing activity by inhib.iting the formation and activation of OC in vitro.